Title: "Discovery of Novel Anti-Inflammatory Phytochemicals on Human Cell Platforms" Project Summary Statement The Problems: Complementary and alternative medicine (CAM) is an understudied scientific discipline. It is unappreciated that many biologically active and therapeutically beneficial compounds are derived from botanicals. Moreover, the scientific methods of high-throughput screening (HTS) for drug discovery and drug toxicity with natural product-derived or synthetic small molecules have become entirely based on speed and volume of small molecules examined. More often than not, a heterologous cell that is non-human and is lacking in in vivo-like properties is the chosen cellular platform;more often than not, synthetic small molecules are chosen to develop forward through medicinal chemistry out of expedience. Together, current HTS approaches have resulted in challenged drug discovery processes, pipelines, and programs across "pharma" that are facing significant "slowdowns." Coupled with that, botanically-derived drug toxicity assessment is expensive and is in need of new and appropriate in vitro human cell models from essential tissues to triage hit- to-lead natural product-derived compounds to the essential few, saving in the time, cost and animal lives. The Solution: DiscoveryBioMed, Inc. (DBM) is an emerging human cell-based drug discovery and drug toxicity company that performs its own R&D programs. DBM also provides this expertise and infrastructure as a high content service for academic and commercial clients. For this NCCAM SBIR Catalyst program, DBM is leading a symbiotic commercial partnership with ChromaDex, Inc. (CDex), a botanically-derived natural products company, to breathe new life into the arena of natural product-based drug discovery and drug toxicity programs. This partnership is formed around the core scientific principle that primary and immortalized human cells are essential and ideal platforms that will drive a new and innovative renaissance in natural product- derived and CAM-friendly basic, applied, translational, drug discovery and drug toxicity sciences. This Catalyst application speaks to specific interests of the NCCAM that seek to discover "the anti-inflammatory biological effects of complex botanicals with a systems biology approach" by finding potent extracts and purified small molecules that protect human skin, respiratory, and kidney cells from harmful irritants and pathogens that cause a myriad of diseases, disorders and syndromes in the human patient. There remains an enormous unmet need to find anti-inflammatories friendly to these different tissues without unwanted side effects. DBM will utilize a deep panel of immortalized human cells derived from skin (keratinocytes), respiratory system (nasal, airway and lung epithelial cells) and renal system (proximal tubule, cortical, and medullary kidney epithelial cells) as platforms for this CAM research. DBM will utilize parental and fractionated extracts derived from botanicals and purified small molecule phytochemicals from plant extracts provided by CDex. Anti- inflammatory extracts and small molecules that protect skin, lung and kidney will be brought forward for potential development. Toxic extracts and small molecules will be assessed, documented, and avoided. The Approach: DBM will assess the effects of botanically-derived extracts and small molecules with high- throughput light-based bioassays of a secreted inflammatory mediator, interleukin-8 (IL-8) that derives from nuclear factor kappa B (NFkappaB) activation, a central immune and inflammatory transcription factor. DBM will seek to solidify hit extracts and small molecules as global lead anti-inflammatory compounds by assessing their ability to also inhibit the secretion of a newly appreciated and important human cytokine, thymic stromal lymphopoietin (TSLP), a lymphocyte chemoattractant and dendritic cell activator that is also upregulated by NFkappaB action. DBM studies its immortal human skin, lung and kidney epithelial cells on extracellular matrix (ECM)-coated microtiter plates. DBM studies any potential cytotoxicity using a light-based CellTiter GLO bioassay that is also HTS- and microtiter plate-friendly. DBM and ChromaDex will document botanically- derived raw extracts, extract fractions, and purified small molecules for 'cytoprotection'versus 'cytotoxicity.'The Bottom Line: We hypothesize that this commercial-driven partnership will be successful in the identification of botanicals with anti-inflammatory potential and will inform the two companies and the scientific and pharma communities at large about the potential of CAM medicines for immune and inflammatory disorders. PUBLIC HEALTH RELEVANCE: Public Health Relevance Statement This partnership between a human cell-based drug discovery company, Discovery BioMed, Inc., and a botanical-derived natural products company, ChromaDex, Inc. seeks to study the biological effects of phytochemical extracts and small molecules as anti-inflammatory substances for any and all diseases and disorders that cause inflammation in the skin, the respiratory system and the renal system. Together, these companies seek to discover and develop botanically-derived therapeutics in a systems biology approach across immortal human skin, lung and kidney cells to determine which plant-derived materials are safe and anti-inflammatory and which may be toxic to the human body. This DBM-led partnership seeks to lead a new renaissance in complementary and alternative medicine (CAM) research for new global anti-inflammatory drugs with this long-term and closely held scientific and commercial vision.